Characterization of METTL3/14-mediated m6A modification in human transcriptome using Nanopore direct RNA sequencing.

Nanopore sequencing reveals a significant reduction in m6A modifications across human transcripts following METTL3/14 depletion, highlighting their distinct roles in RNA regulation.

• Why it matters: Understanding m6A modifications is crucial because they influence RNA metabolism and gene expression, yet current detection methods lack accuracy and reproducibility, limiting insights into their biological functions.
• What they did: The study employed Oxford Nanopore Technologies' direct RNA sequencing with two chemistries and models (m6Anet and Dorado) to profile m6A at single-nucleotide resolution in human cells, validating findings with orthogonal assays and gene knockdowns.
• The result: Findings demonstrate a global decrease in m6A sites and stoichiometry, especially in highly modified genes, with METTL3 and METTL14 showing non-redundant, transcript-specific effects, advancing RNA modification detection and understanding of methyltransferase functions.